BIOB10H3 Lecture Notes - Lecture 6: Fluorescence Microscope, Live Cell Imaging, Fluorophore
12 Sep 2016
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How do we make antibodies for use in the lab: inject human protein into lab animal. Two types of immunofluorescence: direct directly attached flouresceinated antibody, indirect indirectly tagging the fluoresceinateed anti-lg antibodies are proteins possible to generate antibodies against antibodies. Why would you use this? economics: antibodies may be scarce. Gfp: green fluorescent protein from the jellyfish aequorea victoria, recombinant dna is made by attaching the gene then codes for. Gfp with the gene of the protein of interest: this (foreign) dna is introduced into cells in culture. Transfection: dna rna protein, cell will synthesize a chimeric protein with gfp fused to the protein of interest, we can now visualize the protein by using the wavelength of light that excites gfp. Gfp fusion proteins: can look at live" cells and follow protein dynamics, various variants of gfp have now been engineered and glow in different colors.
